四君子汤多糖分离纯化对IEC-6细胞迁移药效活性的影响  

IEC-6 Cell Migration Ability Changes is Impacted by Polysaccharides Separated and Purified from Sijunzi Decoction

作　　者：邓娇[1] 李茹柳[1] 蔡佳仲[1] 涂小华[1] 陈蔚文[1] 

机构地区：[1]广州中医药大学脾胃研究所,广州510006

出　　处：《世界科学技术-中医药现代化》2016年第4期600-606,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委面上项目(81173254):益气健脾中药对小肠上皮细胞迁移过程多胺信号通路中钙离子调控的研究;负责人:李茹柳;广东省中医药局建设中医药强省科研项目(20151190):基于多胺信号通路探讨四君子汤多糖促进小肠上皮细胞迁移的作用机理;负责人:蔡佳仲

摘　　要：目的:本研究对四君子汤多糖进行提取和分离纯化,观察不同多糖样品对小肠上皮细胞(IEC?6)迁移的影响,从而进行多糖纯化过程的药效活性追踪,为探讨四君子汤多糖药效物质基础提供参考。方法:1四君子汤水提醇沉得到四君子汤多糖1;经Sevag法去蛋白得到四君子汤多糖2;经DEAE?52纤维素柱层析并超纯水洗脱得到四君子汤多糖3a;经Sephadex G?100葡聚糖凝胶柱层析并超纯水洗脱得到四君子汤多糖4a和多糖4b;并以苯酚?硫酸法检测各多糖样品糖含量(以葡萄糖计)。2凝胶渗透色谱法(GPC)检测四君子汤多糖3a、多糖4a和多糖4b纯度。3在IEC?6细胞迁移模型上,观察四君子汤多糖不同样品促进细胞迁移的药效活性。结果:4种多糖样品(四君子汤多糖1、2、3a、4b)糖含量分别为71.35%、75.45%、81.00%和98.50%,其促进细胞迁移的最低有效剂量分别为200、100、50、30 mg·L?1;表明纯化步骤最多的多糖4b糖含量最高、药效最优,且GPC法测得其为均一性多糖组分。结论:四君子汤多糖经不同纯化步骤后,糖含量和纯度提高,药效活性随之增强,研究结果为探讨四君子汤促进细胞迁移的药效物质基础研究提供参考。This study aimed to observe the effects of different polysaccharide samples extracted, separated and purified from Sijunzi Decoction（SJZD） on migration ability of the small intestinal epithelial cells（IEC-6）, and track the pharmacodynamic activities of polysaccharide in the purification process, in hoping of providing some references for the pharmacodynamic material research of SJZD. In this study, firstly, the pharmaceutical chemicals of the SJZD were extracted by different purification methods： polysaccharide 1 was separated by hot reflux and ethanol precipitation; polysaccharide 2 was obtained through removing protein by Sevag method; polysaccharide 3a was extracted using DEAE-52 fiber column and ultra pure water elution; while polysaccharide 4a and polysaccharide 4b was purified using Sephadex G-100 gel column chromatography through ultra pure water elution. Thus, glucose content of each sample was detected by phenol-sulfuric acid method. Secondly, the purity degrees of the four polysaccharides were tested by gel permeation chromatography（GPC）. At last, effects of the various polysaccharide samples on the migration of IEC-6 cells were observed by the cell migration model. As a result, the glucose contents of the four polysaccharide samples（polysaccharide 1, 2, 3a, 4b） were 71.35%, 75.45%, 81.0% and 98.5%, respectively; and the lowest effective doses of cell migration ability were 200, 100, 50 and 30 mg·L-1, respectively. These results indicated that polysaccharide 4b was a sort of homogeneous polysaccharide with the highest glucose level and the optimal pharmacodynamics function through the most critical purification steps. It was concluded that the most critical purification method was efficiency in increasing the content and purity of glucose from SJZD and enhancing the pharmacodynamic activities of polysaccharide. All the results provided evidences for exploring pharmacodynamic materials of SJZD in improving cell migration abilities.

关 键 词：四君子汤多糖 分离纯化 IEC-6细胞 细胞迁移 

分 类 号：R289[医药卫生—方剂学]